Fig. 3

Fluorescence polarization assay demonstrating M36 binding to p32. a 5-FAM labeled cyclic LyP-1 peptide was incubated with increasing concentrations of recombinant p32 for 2 h. Increase in fluorescence polarization (mFP) indicates binding to the peptide. b 100 nM p32 was pre-incubated 1 h with increasing concentrations of unlabelled LyP-1 peptide in a competition assay before incubation with 0.5 nM labeled cyclic LyP-1 peptide. c 100 nM p32 was incubated with increasing concentrations of M36 for 30 min and then incubated with 5-FAM labeled cyclic LyP-1 peptide for 2 h. Direct binding of the hit compound M36 to p32 inhibits peptide binding resulting in decreased fluorescence polarization (mFP). Error bars represent standard deviation